LOCUS       sequence-157189-        5420 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Localization: Protein Expression Vector , Excitation: 462, Emission:
            492.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5420)
  AUTHORS   Ai HW, Henderson JN, Remington SJ, Campbell RE
  TITLE     Directed evolution of a monomeric, bright and photostable version of
            Clavularia cyan fluorescent protein: structural characterization and
            applications in fluorescence imaging.
  JOURNAL   Biochem J. 2006 Dec 15. 400(3):531-40.
  PUBMED    16859491
REFERENCE   2  (bases 1 to 5420)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biochem J. 
            2006 Dec 15. 400(3):531-40."
COMMENT     Sequence Label: mTFP1-pBAD
FEATURES             Location/Qualifiers
     source          1..5420
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    5..145
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(71..90)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     231..253
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     391..410
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(590..1468)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1495..1779
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1702..1721
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             1806..1828
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1848..1865
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1869..1901
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             1914..1934
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1941..2651
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     primer_bind     complement(2697..2714)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2867..2953
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3045..3072
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3091..3182
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3183..3251,3252..4043)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3401..3420)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4085..4540
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4172..4191)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4382..4403
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      4651..5239
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5140..5159
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5393..5410
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatctggt
       61 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
      121 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
      181 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
      241 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagaagg agatggcgcc
      301 caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag
      361 cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac
      421 cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga tctgctcatg
      481 tttgacagct tatcatcgat gcataatgtg cctgtcaaat ggacgaagca gggattctgc
      541 aaaccctatg ctactccgtc aagccgtcaa ttgtctgatt cgttaccaat tatgacaact
      601 tgacggctac atcattcact ttttcttcac aaccggcacg gaactcgctc gggctggccc
      661 cggtgcattt tttaaatacc cgcgagaaat agagttgatc gtcaaaacca acattgcgac
      721 cgacggtggc gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt
      781 ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg gcggaaaaga tgtgacagac
      841 gcgacggcga caagcaaaca tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca
      901 ggtgatcgct gatgtactga caagcctcgc gtacccgatt atccatcggt ggatggagcg
      961 actcgttaat cgcttccatg cgccgcagta acaattgctc aagcagattt atcgccagca
     1021 gctccgaata gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga
     1081 aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt attggcaaat attgacggcc
     1141 agttaagcca ttcatgccag taggcgcgcg gacgaaagta aacccactgg tgataccatt
     1201 cgcgagcctc cggatgacga ccgtagtgat gaatctctcc tggcgggaac agcaaaatat
     1261 cacccggtcg gcaaacaaat tctcgtccct gatttttcac caccccctga ccgcgaatgg
     1321 tgagattgag aatataacct ttcattccca gcggtcggtc gataaaaaaa tcgagataac
     1381 cgttggcctc aatcggcgtt aaacccgcca ccagatgggc attaaacgag tatcccggca
     1441 gcaggggatc attttgcgct tcagccatac ttttcatact cccgccattc agagaagaaa
     1501 ccaattgtcc atattgcatc agacattgcc gtcactgcgt cttttactgg ctcttctcgc
     1561 taaccaaacc ggtaaccccg cttattaaaa gcattctgta acaaagcggg accaaagcca
     1621 tgacaaaaac gcgtaacaaa agtgtctata atcacggcag aaaagtccac attgattatt
     1681 tgcacggcgt cacactttgc tatgccatag catttttatc cataagatta gcggatacta
     1741 cctgacgctt tttatcgcaa ctctctactg tttctccata cccgtttttt tgggctagaa
     1801 ataattttgt ttaactttaa gaaggagata tacatatgcg gggttctcat catcatcatc
     1861 atcatggtat ggctagcatg actggtggac agcaaatggg tcgggatctg tacgagaacc
     1921 tgtacttcca gggctcgagc atggtgagca agggcgagga gaccacaatg ggcgtaatca
     1981 agcccgacat gaagatcaag ctgaagatgg agggcaacgt gaatggccac gccttcgtga
     2041 tcgagggcga gggcgagggc aagccctacg acggcaccaa caccatcaac ctggaggtga
     2101 aggagggagc ccccctgccc ttctcctacg acattctgac caccgcgttc gcctacggca
     2161 acagggcctt caccaagtac cccgacgaca tccccaacta cttcaagcag tccttccccg
     2221 agggctactc ttgggagcgc accatgacct tcgaggacaa gggcatcgtg aaggtgaagt
     2281 ccgacatctc catggaggag gactccttca tctacgagat acacctcaag ggcgagaact
     2341 tcccccccaa cggccccgtg atgcagaaga agaccaccgg ctgggacgcc tccaccgaga
     2401 ggatgtacgt gcgcgacggc gtgctgaagg gcgacgtcaa gcacaagctg ctgctggagg
     2461 gcggcggcca ccaccgcgtt gacttcaaga ccatctacag ggccaagaag gcggtgaagc
     2521 tgcccgacta tcactttgtg gaccaccgca tcgagatcct gaaccacgac aaggactaca
     2581 acaaggtgac cgtttacgag agcgccgtgg cccgcaactc caccgacggc atggacgagc
     2641 tgtacaagta agaattcgaa gcttggctgt tttggcggat gagagaagat tttcagcctg
     2701 atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt
     2761 agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat
     2821 ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa
     2881 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
     2941 gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg
     3001 gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac
     3061 ggatggcctt tttgcgtttc tacaaactct tttgtttatt tttctaaata cattcaaata
     3121 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3181 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     3241 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3301 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     3361 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     3421 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     3481 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     3541 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     3601 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     3661 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     3721 tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     3781 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     3841 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     3901 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     3961 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4021 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     4081 atttacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4141 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4201 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4261 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
     4321 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4381 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat
     4441 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4501 tttaacgcga attttaacaa aatattaacg tttacaattt aaaaggatct aggtgaagat
     4561 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4621 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4681 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4741 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     4801 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4861 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4921 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4981 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5041 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5101 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5161 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5221 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5281 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     5341 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     5401 agtgagcgag gaagcggaag
//
